Biochemical sensing array, preparation method, detection test paper and detection kit

Abstract

The invention relates to a biochemical sensor array, a preparation method, detection test paper and a detection kit. The preparation method comprises the following steps: providing a substrate; forming a flexible layer on the second surface in a microelectronic printing mode, wherein the flexible layer at least covers the through hole; a conductive layer is formed on the first face in a microelectronic printing mode, the conductive layer comprises a plurality of discrete conductive blocks, and the conductive blocks cover the set of through holes, the first point positions and the second point positions; a sensitive layer is formed on the side, away from the substrate, of the flexible layer, the sensitive layer comprises a plurality of discrete thin film blocks, and the thin film blocks and the through holes are at least partially overlapped. Therefore, the flexible layer and the conductive layer are prepared in a microelectronic printing mode, the film layer thickness of the flexible layer and the conductive layer is accurately controlled, and the uniformity and repeatability of the biosensor are improved; the prepared biochemical sensor array comprises a plurality of biochemical sensors, batch detection can be carried out, and compared with a single biochemical sensor, the detection efficiency is remarkably improved.

Description

Technical Field

[0001] The present disclosure relates to the technical field of biosensors, and particularly to a biochemical sensing array, a preparation method, a test strip and a test kit. Background Art

[0002] A surface stress biosensor is a type of biosensor, which can be applied to fields such as clinical medical diagnosis, biochemical detection, and new drug discovery. The detection principle of the surface stress biosensor is that specific probe modification of the sensor can achieve qualitative analysis of the biological molecules to be detected; after the biological molecules to be detected are combined with their corresponding probes, the sensitive component of the biosensor will produce a small deformation, and the stress generated by the combination of different concentrations of the biological molecules to be detected and the probes is different. By detecting the electrical signals of different deformations, quantitative analysis of the concentration of the biological molecules to be detected can be achieved.

[0003] In the related art, each film layer of the sensor is usually prepared by a spin coating method, which is difficult to accurately control the thickness of the film layer, resulting in poor uniformity and repeatability of the prepared biosensor. Summary of the Invention

[0004] In order to solve the above technical problems, the present disclosure provides a biochemical sensing array, a preparation method, a test strip and a test kit.

[0005] In a first aspect, the present disclosure provides a preparation method of a biochemical sensing array, including:

[0006] Providing a substrate; the substrate includes a first surface and a second surface arranged oppositely, the first surface is provided with a plurality of through holes arranged in an array, a first trace and a plurality of second traces; the second traces correspond to the through holes one by one, a first point and a second point are provided on the circumferential of the through hole, the first trace is electrically connected to all the first points, and the second traces are electrically connected to the second points one by one;

[0007] Forming a flexible layer on the second surface by a microelectronic printing method; the flexible layer at least covers the through holes;

[0008] Forming a conductive layer on the first surface by a microelectronic printing method; the conductive layer includes a plurality of discrete conductive blocks, and the conductive blocks cover a group of the through holes, the first points and the second points;

[0009] Forming a sensitive layer on the side of the flexible layer facing away from the substrate; the sensitive layer includes a plurality of discrete thin film blocks, and the thin film blocks at least partially overlap with the through holes.

[0010] In a second aspect, the present disclosure further provides a biochemical sensing array, and the biochemical sensing array is formed based on the preparation method of any one of the above biochemical sensing arrays.

[0011] In a third aspect, the present disclosure also provides a test strip, comprising any one of the above biochemical sensing arrays.

[0012] In a fourth aspect, the present disclosure also provides a test kit, comprising the above test strip.

[0013] The technical solution provided by the present disclosure has the following advantages compared with the prior art:

[0014] The biochemical sensing array, preparation method, test strip and test kit provided by the present disclosure, the preparation method comprising: providing a substrate; the substrate includes a first surface and a second surface arranged opposite to each other, the first surface is provided with a plurality of through holes arranged in an array, a first trace and a plurality of second traces; the second traces correspond to the through holes one by one, the circumferences of the through holes are provided with a first point and a second point, the first trace is electrically connected to all the first points, and the second traces are electrically connected to the second points one by one; forming a flexible layer on the second surface in a microelectronic printing manner; the flexible layer at least covers the through holes; forming a conductive layer on the first surface in a microelectronic printing manner; the conductive layer includes a plurality of discrete conductive blocks, and the conductive blocks cover a group of through holes, the first points and the second points; forming a sensitive layer on the side of the flexible layer away from the substrate; the sensitive layer includes a plurality of discrete thin film blocks, and the thin film blocks at least partially overlap with the through holes. Thus, the flexible layer and the conductive layer are prepared by microelectronic printing, and the film thicknesses of the flexible layer and the conductive layer are accurately controlled, which is beneficial to improving the uniformity and repeatability of the biochemical sensor; the prepared biochemical sensing array includes a plurality of biochemical sensors and can perform batch detection. Compared with a single biochemical sensor, the detection efficiency is significantly improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The drawings herein are incorporated into the specification and form a part of the specification, showing embodiments consistent with the present disclosure, and are used together with the specification to explain the principles of the present disclosure.

[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present disclosure or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, for those of ordinary skill in the art, other drawings can also be obtained based on these drawings without creative efforts.

[0017] Figure 1 It is a schematic flow chart of a method for preparing a biochemical sensing array provided by an embodiment of the present disclosure;

[0018] Figures 2-6 It is a schematic structural diagram corresponding to each step in the method for preparing a biochemical sensing array provided by an embodiment of the present disclosure;

[0019] Figure 7 It is a schematic structural diagram of a substrate provided by an embodiment of the present disclosure;

[0020] Figure 8 Schematic diagram of the detection result of the biochemical sensing array provided by the embodiment of the present disclosure. Detailed implementation manners

[0021] In order to be able to more clearly understand the above-mentioned objects, features, and advantages of the present disclosure, the solutions of the present disclosure will be further described below. It should be noted that, without conflict, the embodiments of the present disclosure and the features in the embodiments can be combined with each other.

[0022] Many specific details are set forth in the following description in order to fully understand the present disclosure, but the present disclosure may also be implemented in other ways different from those described herein; obviously, the embodiments in the specification are only a part of the embodiments of the present disclosure, rather than all the embodiments.

[0023] Figure 1 Schematic flow chart of a preparation method of a biochemical sensing array provided by the embodiment of the present disclosure. Referring to Figure 1 , the preparation method of the biochemical sensing array includes the following steps:

[0024] S110. Provide a substrate.

[0025] Combined with Figures 2-3 , the substrate 10 includes a first surface S1 and a second surface S2 which are oppositely arranged. The first surface S1 is provided with a plurality of through holes 11 arranged in an array, a first trace 12, and a plurality of second traces 13; the second traces 13 correspond to the through holes 11 one by one. A first point 14 and a second point 15 are provided on the circumference of the through hole 11. The first trace 12 is electrically connected to all the first points 14, and the second traces 13 are electrically connected to the second points 15 one by one.

[0026] Exemplarily, the first surface S1 is the front surface of the substrate 10, and the second surface S2 is the back surface of the substrate 10.

[0027] Exemplarily, as Figure 2 shown, the first surface S1 of the substrate 10 is provided with 25 through holes 11, 1 first trace 12, and 25 second traces 13; 1 first point 14 and 1 second point 15 are provided on the circumference of each through hole 11. The first trace 12 is electrically connected to all the first points 14, and the second traces 13 are electrically connected to the second points 15 one by one.

[0028] In some embodiments, as Figure 2 shown, the first surface S1 of the substrate 10 is further provided with a signal interface 16. The signal interface 16 includes a first signal interface 161 and a second signal interface 162 arranged in sequence along the first direction X. The first signal interface 161 is electrically connected to the first trace 12, and the second signal interface 162 is electrically connected to the second traces 13 one by one.

[0029] Among them, the substrate 10 includes a flexible substrate, such as a Flexible Printed Circuit (FPC).

[0030] S120: Form a flexible layer on the second surface in a microelectronic printing manner.

[0031] Combined with Figure 4 , the flexible layer 20 covers at least the through holes 11.

[0032] Exemplarily, the flexible layer 20 only covers the through holes 11.

[0033] Exemplarily, as Figure 4 shown, the flexible layer 20 covers the through holes and the surface of the substrate 10 around the circumference of the through holes 11 (i.e., part of the second surface S2).

[0034] Exemplarily, the flexible layer 20 covers the through holes 11 and the entire second surface S2 of the substrate 10.

[0035] In this step, the second surface S2 of the substrate 10 is the printing surface. Place the substrate 10 at a preset position, and set the printing position of the flexible layer 20 based on the printing coordinate system. The printing position of the flexible layer 20 includes a printing start position and a printing end position.

[0036] The embodiments of the present application do not limit the material of the flexible layer 20, and all flexible materials known to those skilled in the art can be used, such as polydimethylsiloxane (PDMS), polyimide, polyethylene, ethylene-vinyl acetate copolymer, polyvinyl alcohol, polyethylene terephthalate, and polymethyl methacrylate.

[0037] S130: Form a conductive layer on the first surface in a microelectronic printing manner.

[0038] Combined with Figure 5 , the conductive layer 30 includes a plurality of discrete conductive blocks 31. The conductive blocks 31 cover a group of through holes 11, the first point 14, and the second point 15. Exemplarily, as Figure 5 shown, 25 through holes 11 are provided on the substrate 10. One through hole 11 and the first point 14 and the second point 15 arranged around it form a group, and there are 25 groups in total. Then the conductive layer 30 includes 25 discrete conductive blocks 31, and each conductive block 31 covers a group of through holes 11, the first point 14, and the second point 15.

[0039] In this step, the first surface S1 of the substrate 10 is the printing surface. The substrate 10 is placed at a preset position, and the printing position of the conductive block 31 is set based on the printing coordinate system, so that the conductive block 31 covers the upper surfaces of each group of through holes 11, the first point 14, and the second point 15. The formed conductive block 31 contacts the first point 14 and the second point 15 to form an electrical connection, thereby conducting the first trace 12 and the second trace 13 of this group of sensors.

[0040] The present disclosure embodiment does not limit the preparation material of the conductive layer 30, including but not limited to silver nanowires, carbon nanotubes, conductive silver paste, graphene, and two-dimensional inorganic compound MXene.

[0041] S140. Form a sensitive layer on the side of the flexible layer facing away from the substrate.

[0042] Combined Figure 6 , the sensitive layer 40 includes a plurality of discrete thin film blocks 41, and the thin film blocks 41 at least partially overlap with the through holes 11. Exemplarily, as Figure 6 shown, 25 through holes 11 are provided on the substrate 10, the sensitive layer 40 includes 25 discrete thin film blocks 41, and the thin film blocks 41 overlap with the through holes 11.

[0043] In this step, the mask plate is overlapped with the second surface S2 of the substrate. The mask plate is also provided with through holes arranged in an array, and the through holes on the mask plate at least partially overlap with the through holes 11 on the substrate 10; then a sensitive layer 40 is formed on the side of the mask plate facing away from the substrate 10. After removing the mask plate, discrete thin film blocks 41 are formed on the side of the flexible layer 20 facing away from the through holes 11. In some embodiments, the substrate 10 in the S110 step is used as the mask plate, and the substrate 10 is covered on another substrate 10 having a flexible layer 20, and the through holes 11 of the two substrates 10 overlap.

[0044] In this embodiment, microelectronic printing has the advantage of adjustable printing parameters, can accurately control the film layer thickness, realize the standardized preparation of the film layer, and thus realize the standardized preparation of biochemical sensors. The obtained biochemical sensing array belongs to a surface stress biochemical sensor, which includes a plurality of biochemical sensors arranged in an array. Each through hole corresponds to a biochemical sensor. The biochemical sensors have good uniformity and repeatability, can be used for batch detection, and significantly improve the detection accuracy and detection efficiency.

[0045] Using a microelectronic printer to print the flexible layer 20 and the conductive layer 30 (or the conductive block 31), the control accuracy of the printed film layer thickness can reach the nanometer level.

[0046] In some embodiments, the steps of S130 and S140 can also be exchanged, first preparing the sensitive layer 40, and then preparing the conductive layer 30.

[0047] It should be noted thatFigures 2-6 Exemplarily, it is illustrated that the biochemical sensing array includes 25 biochemical sensors, which are arranged in a 5×5 array. The shapes of the first position 14 and the second position 15 are both square, but this does not limit the preparation method of the biochemical sensing array provided by the embodiments of the present application. In other embodiments, fewer or more biochemical sensors may be provided on the biochemical sensing array, such as 32 or 20. The first position 14 and the second position 15 may also be set to other shapes, such as rectangular, circular or triangular, which are not limited herein.

[0048] The preparation method of the biochemical sensing array provided by the embodiments of the present disclosure includes: providing a substrate 10; the substrate 10 includes a first surface S1 and a second surface S2 which are oppositely arranged. The first surface S1 is provided with a plurality of through holes 11 arranged in an array, a first trace 12 and a plurality of second traces 13; the second traces 13 correspond to the through holes 11 one by one. The circumferences of the through holes 11 are provided with a first position 14 and a second position 15. The first trace 12 is electrically connected to all the first positions 14, and the second traces 13 are electrically connected to the second positions 15 one by one; forming a flexible layer 20 on the second surface in a microelectronic printing manner; the flexible layer 20 at least covers the through holes 11; forming a conductive layer 30 on the first surface in a microelectronic printing manner; the conductive layer 30 includes a plurality of discrete conductive blocks 31, and the conductive blocks 31 cover a group of through holes 11, the first position 14 and the second position 15; forming a sensitive layer 40 on the side of the flexible layer 20 away from the substrate 10; the sensitive layer 40 includes a plurality of discrete thin film blocks 41, and the thin film blocks 41 at least partially overlap with the through holes 11. Thus, the flexible layer 20 and the conductive layer 30 are prepared by microelectronic printing, and the film thicknesses of the flexible layer 20 and the conductive layer 30 are accurately controlled, which is beneficial to improving the uniformity and repeatability of the biochemical sensors; the prepared biochemical sensing array includes a plurality of biochemical sensors, and can perform batch detection. Compared with a single biochemical sensor, the detection efficiency is significantly improved.

[0049] In some embodiments, the flexible layer 20 includes a polydimethylsiloxane thin film layer; "forming a flexible layer on the second surface in a microelectronic printing manner" includes the following steps:

[0050] Mixing a prepolymer and a curing agent evenly at a mass ratio of M:1 to obtain a mixed glue, and evacuating the mixed glue to remove air bubbles; wherein, 8≤M≤10;

[0051] Using the mixed glue as a raw material, printing a polydimethylsiloxane thin film layer by a dispensing method.

[0052] In this embodiment, before printing the polydimethylsiloxane thin film layer, a mixed glue needs to be prepared; the prepared mixed glue is poured into the raw material box of the printer, the second surface S2 of the substrate 10 is placed on the stage of the printing device, and the first printing parameters are set; after printing, wait for the polydimethylsiloxane thin film layer to solidify, and the substrate 10 covered with the polydimethylsiloxane thin film layer is obtained.

[0053] Among them, the first printing parameters at least include the first printing speed and the first printing air pressure.

[0054] In some embodiments, "printing the polydimethylsiloxane thin film layer by the dispensing method" includes the following steps:

[0055] Based on the first printing speed and the first printing air pressure, print the polydimethylsiloxane thin film layer by the dispensing method;

[0056] Among them, the first printing speed is: 5 - 15 mm / s, and the first printing air pressure is: 30 - 90 KPa.

[0057] Among them, the thickness of the printed polydimethylsiloxane thin film layer is negatively correlated with the first printing speed. The faster the first printing speed, the smaller the thickness of the polydimethylsiloxane thin film layer; the slower the first printing speed, the larger the thickness of the polydimethylsiloxane thin film layer. The thickness of the printed polydimethylsiloxane thin film layer is positively correlated with the first printing air pressure. The larger the first printing air pressure, the larger the thickness of the polydimethylsiloxane thin film layer; the smaller the first printing air pressure, the smaller the thickness of the polydimethylsiloxane thin film layer.

[0058] In this embodiment, the first printing speed is set to: 5 - 15 mm / s, and the first printing air pressure is set to: 30 - 90 KPa, so as to control the polydimethylsiloxane thin film layer within an appropriate thickness range, taking into account both sensitivity and impact resistance. If the first printing speed is set greater than 15 mm / s and / or the first printing air pressure is less than 30 KPa, the thickness of the polydimethylsiloxane thin film layer is too thin, and it is easy to break and damage. If the first printing speed is set less than 5 mm / s and / or the first printing air pressure is greater than 90 KPa, the thickness of the polydimethylsiloxane thin film layer is too thick, and when detecting the biological molecules to be measured, the deformation amount of the sensor decreases, reducing the sensitivity of the sensor.

[0059] Exemplarily, the first printing speed is set to 6 mm / s, or the first printing speed is set to 8 mm / s, or the first printing speed is set to 12 mm / s.

[0060] Exemplarily, the first printing air pressure is set to 90 KPa, or the first printing air pressure is set to 80 KPa.

[0061] In some embodiments, after "printing the polydimethylsiloxane thin film layer by dispensing", the preparation method further includes the following steps:

[0062] Heat the substrate at a first heating temperature until the polydimethylsiloxane thin film layer solidifies.

[0063] In this embodiment, the freshly printed polydimethylsiloxane thin film layer still has a certain viscosity. By heating, its solidification is accelerated, thereby shortening the preparation time and also avoiding damaging the polydimethylsiloxane thin film layer in subsequent steps.

[0064] In this embodiment, the first heating temperature is set to 60 - 80 °C. On the premise of not damaging the structure of the polydimethylsiloxane thin film layer, the solidification time of the polydimethylsiloxane thin film layer is shortened as much as possible. If the first heating temperature is too low (less than 60 °C), the solidification rate of the polydimethylsiloxane thin film layer is slow and the waiting time is long; if the first heating temperature is too high (greater than 80 °C), it exceeds the melting point of the unfrozen polydimethylsiloxane, causing the polydimethylsiloxane thin film layer to bubble and damaging the film layer structure.

[0065] Exemplarily, the first heating temperature is 70 °C, or the first heating temperature is 75 °C.

[0066] It should be noted that all devices or equipment with heating functions known to those skilled in the art can be used, such as a heating table or a drying oven, which is not limited here. There is a certain difference between the set temperature and the actual temperature of the heating device. The set temperature is a single temperature value, and the actual temperature will fluctuate within a preset range of the set temperature. For example, when the set temperature is 80 °C, the actual temperature is 80 ± 5 °C. Another example, when the set temperature is 60 °C, the actual temperature is 60 ± 3 °C.

[0067] Exemplarily, the first heating temperature is one of 65 °C, 70 °C, and 75 °C.

[0068] In some embodiments, the conductive layer 30 includes a silver paste layer; "forming the conductive layer on the first side by microelectronic printing" includes the following steps:

[0069] Using conductive silver paste as the raw material, print conductive blocks by dispensing.

[0070] In this embodiment, pour the conductive silver paste into the raw material box of the printer, place the first side S1 of the substrate 10 at the stage of the printing device, set the second printing parameters; after printing, wait for the silver paste layer to solidify, and then the substrate 10 covered with a plurality of silver paste conductive blocks is obtained. The silver paste conductive blocks prepared in this embodiment have good electrical conductivity. Each silver paste conductive block contacts and is electrically connected to a first point 14 and a second point in the same group, thereby conducting the first trace 12 and the second trace 13 of the sensors in this group.

[0071] The second printing parameter includes at least a second printing speed and a second printing air pressure.

[0072] In some embodiments, "printing the conductive block by the dispensing method" includes the following steps:

[0073] Based on the second printing speed and the second printing air pressure, print the conductive block by the dispensing method;

[0074] Among them, the second printing speed is: 4 - 15 mm / s, and the second printing air pressure is: 70 - 100 KPa.

[0075] Among them, the thickness of the printed conductive block 31 is negatively correlated with the second printing speed. The faster the second printing speed, the smaller the thickness of the conductive block 31, and the slower the first printing speed, the larger the thickness of the conductive block 31. The thickness of the printed conductive block 31 is positively correlated with the second printing air pressure. The larger the second printing air pressure, the larger the thickness of the conductive block 31, and the smaller the second printing air pressure, the smaller the thickness of the conductive block 31.

[0076] In this embodiment, the second printing speed is set to: 4 - 15 mm / s, and the second printing air pressure is set to: 70 - 100 KPa, so as to control the thickness of the conductive block within an appropriate range, taking into account both reliability and cost. If the first printing speed is set to be greater than 15 mm / s and / or the first printing air pressure is less than 70 KPa, the thickness of the conductive block 31 will be too thin, resulting in the non - conduction of the first trace 12 and the second trace 13, affecting the reliability of the sensor. If the first printing speed is set to be less than 4 mm / s and / or the first printing air pressure is greater than 100 KPa, the thickness of the conductive block 31 will be too thick, increasing the manufacturing cost.

[0077] Exemplarily, the second printing speed is set to 4 mm / s, or the second printing speed is set to 5 mm / s, or the first printing speed is set to 10 mm / s.

[0078] Exemplarily, the second printing air pressure is set to 80 KPa, or the second printing air pressure is set to 100 KPa.

[0079] In some embodiments, after "printing the conductive block by the dispensing method", the preparation method further includes the following steps:

[0080] Heat the substrate at the second heating temperature until the conductive block solidifies;

[0081] Among them, the second heating temperature is: 80 - 120 °C, and the heating duration is: 30 - 50 min.

[0082] In this embodiment, the just - printed silver paste layer still has a certain fluidity. By heating, its solidification is accelerated, thereby shortening the preparation time and also avoiding the subsequent operations from affecting the uniformity of the film layer thickness and damaging the structure of the conductive block 31.

[0083] The second heating duration is related to the thickness of the silver paste layer and the second heating temperature. On the premise that the thickness of the silver paste layer is fixed, the higher the second heating temperature, the shorter the second heating duration required for the silver paste layer to solidify; the lower the second heating temperature, the longer the second heating duration required for the silver paste layer to solidify. In this embodiment, the second heating temperature is set to 80-120 °C, which not only accelerates the solidification of the silver paste layer, but also has no impact on the already solidified polydimethylsiloxane thin film layer.

[0084] Exemplarily, the second heating temperature is 80 °C, or the second heating temperature is 100 °C, or the second heating temperature is 120 °C.

[0085] Exemplarily, the second heating duration is 30 min, or the second heating duration is 40 min, or the second heating duration is 50 min.

[0086] It should be noted that all devices or equipment with heating functions known to those skilled in the art can be used, such as a heating table or a drying oven, which is not limited here. There is a certain difference between the set temperature and the actual temperature of the heating device. The set temperature is a single temperature value, and the actual temperature will fluctuate within a preset range of the set temperature. For example, when the set temperature is 100 °C, the actual temperature is 100±5 °C. Another example, when the set temperature is 90 °C, the actual temperature is 90±5 °C.

[0087] In some embodiments, the sensitive layer 40 includes a nano-gold layer; the step of "forming the sensitive layer on the side of the flexible substrate facing away from the substrate" includes the following steps:

[0088] Forming a thin film block on the side of the flexible layer facing away from the substrate by magnetron sputtering.

[0089] In this embodiment, the nano-gold layer is prepared by magnetron sputtering, which can precisely control the thickness and uniformity of the nano-gold layer. The prepared nano-gold layer is a thin film with high purity, density, and uniformity, and has a strong bonding force with the flexible layer 20, high mechanical strength. At the same time, it also has advantages such as high sputtering rate, simple operation, low working temperature, low energy consumption, and environmental protection.

[0090] The magnetron sputtering conditions at least include the first sputtering duration.

[0091] Among them, the material of the nano-gold layer is high-purity gold, and the particle size of its single particle reaches the nanoscale. Nano-gold has good biocompatibility and can bind to a variety of biomolecules without affecting biological activity. The nano-gold layer prepared in this embodiment includes a plurality of discrete thin film blocks 41, and the thin film blocks 41 at least partially overlap with the through holes 11.

[0092] In some embodiments, the step of "forming a thin film block on the side of the flexible layer facing away from the substrate by magnetron sputtering" includes the following steps:

[0093] Sputter a thin film block on the side of the flexible layer facing away from the substrate based on the first sputtering duration;

[0094] Among them, the first sputtering duration is: 1 - 2 min.

[0095] In this embodiment, by adjusting the first sputtering duration, the thickness of the nano - gold layer is controlled. The longer the first sputtering duration, the greater the thickness of the nano - gold layer, and the shorter the first sputtering duration, the smaller the thickness of the nano - gold layer. Controlling the first sputtering duration within the range of 1 - 2 min ensures both the binding performance between the nano - gold layer and the antibody and prevents the film from being too thick, resulting in increased costs.

[0096] Exemplarily, the preparation method of the biochemical sensing array includes the following steps:

[0097] (1) Prepare a mixed glue of polydimethylsiloxane. Mix glue A (pre - polymer) and glue B (curing agent) in a mass ratio of 10:1 and stir evenly, and then evacuate to remove air bubbles.

[0098] (2) Print a polydimethylsiloxane thin film layer. After pouring the prepared mixed glue into the raw material box of the micro - electronics printer, place the back of the flexible substrate on the stage, set the printing speed to 8 mm / s, and the air pressure to 90 KPa; after printing is completed, heat the flexible substrate at 80 °C. After the polydimethylsiloxane thin film layer solidifies, a flexible substrate covered with a polydimethylsiloxane thin film layer on the back can be obtained.

[0099] (3) Print a silver paste layer. After pouring the conductive silver paste into the raw material box of the micro - electronics printer, place the front of the flexible substrate on the stage of the printer, set the printing speed to 5 mm / s, and the air pressure to 80 KPa; after printing is completed, heat the flexible substrate at 120 °C for 30 min, and a flexible substrate covered with a conductive layer on the front can be obtained.

[0100] (4) Form a nano - gold layer. Use a magnetron sputtering instrument to sputter a nano - gold layer on the surface of the polydimethylsiloxane thin film to obtain a nano - gold - polydimethylsiloxane (AuNPs - PDMS) composite film.

[0101] Exemplarily, the preparation method of the biochemical sensing array includes the following steps:

[0102] (1) Prepare a mixed glue of polydimethylsiloxane. Mix glue A (pre - polymer) and glue B (curing agent) in a mass ratio of 10:1 and stir evenly, and then evacuate to remove air bubbles.

[0103] (2) Print the polydimethylsiloxane thin film layer. After pouring the prepared mixed glue into the raw material box of the microelectronic printer, place the back side of the flexible substrate on the stage, set the printing speed to 6 mm / s, and the air pressure to 80 KPa; after printing is completed, place the flexible substrate in an 80°C oven for heating. After the polydimethylsiloxane thin film layer solidifies, a flexible substrate with the back side covered by the polydimethylsiloxane thin film layer can be obtained.

[0104] (3) Print the silver paste layer. After pouring the conductive silver paste into the raw material box of the microelectronic printer, place the front side of the flexible substrate on the stage of the printer, set the printing speed to 4 mm / s, and the air pressure to 100 KPa; after printing is completed, place the flexible substrate in a 120°C oven for heating for 30 min, and a flexible substrate with the front side covered by the conductive layer can be obtained.

[0105] (4) Form the nano-gold layer. Use a magnetron sputtering instrument to sputter a nano-gold layer on the surface of the polydimethylsiloxane thin film to obtain a nano-gold-polydimethylsiloxane (AuNPs-PDMS) composite film.

[0106] In some embodiments, the preparation method further includes the following steps:

[0107] Modify the antibody on the side of the film block facing away from the flexible layer;

[0108] Among them, the antibody is used to specifically bind to the biomolecule to be detected.

[0109] In this embodiment, an antibody solution is dropped on each film block 41 of the sensitive layer 40 and maintained for a preset duration, so that antibody molecules are modified onto the film block 41. When using the biochemical sensing array prepared in this embodiment for detection, solutions of different concentrations of the biomolecule to be detected are respectively dropped on different biochemical sensors, so that the biomolecule to be detected specifically binds to the antibody; the films of the flexible layer and the sensitive layer are deformed, so that the distance between the conductive silver pastes in the approximate conductive block becomes larger, the conductive paths are reduced, and the resistance of the sensor increases. The change value of the resistance can be detected by a biological detector to realize the detection of the biomolecule to be detected.

[0110] Exemplarily, the sensitive layer 40 is a nano-gold layer. An antibody solution is dropped on each film block of the nano-gold layer and maintained for a preset duration, so that antibody molecules are modified onto the nano-gold layer; then the biochemical sensing array is rinsed with a buffer solution to inhibit non-specific adsorption.

[0111] The preparation method of the biochemical sensing array provided by the embodiment of the present application uses green and environmentally friendly raw materials, does not involve toxic materials such as fluorescent reagents, does not cause environmental pollution, and conforms to the green development concept of our country. At the same time, quantitative analysis and scientific evaluation are carried out on the specific data of the possible ecological benefits, indicating that it has a great promoting effect on the sustainable development of the economy and the full utilization of resources.

[0112] Based on the above embodiments, the embodiments of the present disclosure further provide a biochemical sensing array, which is formed by the preparation method of any of the above biochemical sensing arrays and has corresponding beneficial effects. The same parts can be understood with reference to the above text and will not be repeated hereinafter.

[0113] In some embodiments, as Figures 2-6 shown, the biochemical sensing array includes a substrate 10, a flexible layer 20, a conductive layer 30, and a sensitive layer 40.

[0114] Among them, the substrate 10 includes a first surface S1 and a second surface S2 arranged opposite to each other. The first surface S1 is provided with a plurality of through holes 11 arranged in an array, a first trace 12, and a plurality of second traces 13; the second traces 13 correspond to the through holes 11 one by one. A first point 14 and a second point 15 are provided on the circumference of the through hole 11. The first trace 12 is electrically connected to all the first points 14, and the second traces 13 are electrically connected to the second points 15 one by one.

[0115] The flexible layer 20 is located on the second surface S2 of the substrate 10 and covers at least the through holes 11.

[0116] The sensitive layer is located on the side of the flexible layer 20 away from the substrate 10. The sensitive layer 40 includes a plurality of discrete thin film blocks 41, and the thin film blocks 41 at least partially overlap with the through holes 11.

[0117] The conductive layer 30 is located on the first surface S1 of the substrate 10. The conductive layer 30 includes a plurality of discrete conductive blocks 31. The conductive blocks 31 cover a group of through holes 11, the first points 14, and the second points 15. The conductive blocks 31 are in contact with the first points 14 and the second points 15 to form an electrical connection, thereby conducting the first trace 11 and the second traces 13 of the group of sensors.

[0118] Among them, the number of biochemical sensors included in the biochemical sensing array is equal to the number of through holes 11. Exemplarily, as Figures 2-6 shown in any figure, the number of through holes 11 is 25, and the biochemical sensing array includes 25 biochemical sensors.

[0119] It should be noted that Figures 2-6 exemplarily illustrates that the biochemical sensing array includes 25 biochemical sensors and is arranged in a 5×5 array manner. The shapes of the first point 14 and the second point 15 are both squares, but it does not constitute a limitation on the biochemical sensing array provided by the embodiments of the present application. In other embodiments, fewer or more biochemical sensors can be provided on the biochemical sensing array, such as 32 or 20. The first point 14 and the second point 15 can also be set to other shapes, such as rectangles, circles, or triangles, which are not limited herein.

[0120] In some embodiments, as Figure 2 shown, a signal interface 16 is further provided on the first surface S1 of the substrate 10. The signal interface 16 includes a first signal interface 161 and a second signal interface 162 arranged in sequence. The first signal interface 161 is electrically connected to the first trace 12, and the second signal interface 162 is electrically connected to the second trace 13 in one-to-one correspondence.

[0121] Among them, the biochemical sensing array includes multiple signal access channels. One signal access channel includes a first signal interface 161, a second signal interface 162, and a biochemical sensor correspondingly connected between the first signal interface 161 and the second signal interface 162.

[0122] When using a biological detector to measure the biochemical sensing array, insert the signal interface 16 of the biochemical sensing array into the card slot of the biological detector, so that the first signal interface 161 and the second signal interface 162 are electrically connected to the detection circuit in the biological detector. The first signal interface 161 serves as a common signal interface, and the second signal interface 162 serves as a detection signal interface to detect the signals of each signal access channel. Exemplarily, the signals of the signal access channel include resistance change values.

[0123] Exemplarily, as Figure 2 shown, 25 through holes 11 are provided on the substrate 10. The through holes 11 are arranged in a 5×5 array, corresponding to 25 groups of first points and second points, 1 first trace 12 and 1 first signal interface 161, 25 second traces 13 and 25 second signal interfaces 162; one end of the first trace 12 is electrically connected to the first signal interface 161, and the other end is electrically connected to all the first points 14; one end of the second trace 13 is electrically connected to the second signal interface 162 in one-to-one correspondence, and the other end is electrically connected to the second point 15 in one-to-one correspondence.

[0124] In some embodiments, the diameter of the through hole is: 2.0 - 5.0 mm; the distance between the centers of adjacent through holes is: 3.0 - 15 mm.

[0125] In some embodiments, the distance between the edge of the first point and the edge of the corresponding through hole is: 0.3 - 1.0 mm; the distance between the edge of the second point and the edge of the corresponding through hole is: 0.3 - 1.0 mm.

[0126] In some embodiments, the line width of the first trace is: 0.2 - 0.5 mm; the line width of the second trace is: 0.2 - 0.5 mm.

[0127] In some embodiments, the line spacing between the first trace and the second trace and between the second traces is: 0.2 - 0.5 mm.

[0128] In some embodiments, the width of the first signal interface is 0.3 - 0.7 mm; the width of the second signal interface is 0.3 - 0.7 mm.

[0129] Exemplarily, as Figure 7 shown, along the first direction X, the length L1 of the substrate 10 is 70 mm, the length L3 of the signal interface is 28.5 mm, the distance L5 between the centers of adjacent through-holes 11 is 13.4 mm, the distance L6 between the edge of the first position 14 and the edge of the corresponding through-hole 11 is 0.5 mm, and the distance L7 between the edge of the second position 15 and the edge of the corresponding through-hole 11 is 0.5 mm; along the second direction Y, the width L2 of the array distribution area of the substrate 10 is 42 mm, and the width L4 of the trailing area is 10 mm; the distance L5 between the centers of adjacent through-holes 11 is 5 mm, and the line distance L8 between the first trace 12 and the second trace 13 and between the second traces 13 is 0.5 mm; the diameter Φ of the through-hole 11 is 3 mm; the line widths of both the first trace and the second trace are 0.3 mm; along the first direction X, the width of the first signal interface 161 is 0.5 mm, and the width of the second signal interface 162 is 0.5 mm.

[0130] Exemplarily, the biochemical sensing array provided by the present disclosure is used to detect Escherichia coli O157:H7, and the specific detection process is as follows:

[0131] (1) Dilute Escherichia coli O157:H7 to 50, 100, 150, 200, and 250 CFU / mL respectively.

[0132] (2) Drop the above-mentioned Escherichia coli O157:H7 with different concentrations onto the biochemical sensor and react for 30 min to enable specific binding between Escherichia coli O157:H7 and the antibody.

[0133] (3) Use a portable in vitro biological detector to measure the resistance value. Combining Figure 5 , the biochemical sensing array has 26 signal interfaces. The first one on the left is the first signal interface (common signal interface), and the remaining 25 are the second signal interfaces (measurement signal interfaces); between each second signal interface and the first signal interface is an impedance test area. If the impedance of the solution in the test area changes, the resistance measured by these two signal interfaces will change.

[0134] (4) The detection results are as Figure 8 shown. The resistance change value has a linear relationship with the concentration of Escherichia coli O157:H7, and the resistance change value increases with the increase of the concentration of Escherichia coli O157:H7. The variance R 2 of the fitting formula is equal to 0.98, indicating that the biochemical sensing array has good uniformity and repeatability.

[0135] Based on the above embodiments, the embodiments of the present disclosure further provide a test strip, which includes any of the above biochemical sensing arrays and has corresponding beneficial effects. To avoid repeated description, it will not be elaborated here.

[0136] Based on the above embodiments, the embodiments of the present disclosure further provide a test kit, which includes the above test strip and has corresponding beneficial effects. To avoid repeated description, it will not be elaborated here.

[0137] In other embodiments, the test kit further includes all constituent structures known to those skilled in the art, such as a housing, a test reagent, a dropper, a seal bag, etc., which are not limited herein.

[0138] It should be noted that in this article, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not expressly listed, or further includes elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "comprising one..." does not exclude the existence of another identical element in the process, method, article or device comprising the element.

[0139] The above are only specific embodiments of the present disclosure, enabling those skilled in the art to understand or implement the present disclosure. Various modifications to these embodiments will be obvious to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present disclosure. Therefore, the present disclosure will not be limited to these embodiments described herein, but will conform to the broadest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a biochemical sensing array, characterized in that, include: providing a substrate; The substrate comprises a first surface and a second surface arranged opposite to each other, the first surface is provided with a plurality of through holes arranged in an array, a first routing line and a plurality of second routing lines; the second routing lines correspond to the through holes one by one, the through holes are provided with first points and second points around the through holes, the first routing line is electrically connected to all the first points, and the second routing lines are electrically connected to the second points one by one; Forming a flexible layer on the second surface by microelectronic printing; the flexible layer at least covers the through hole; A conductive layer is formed on the first surface by microelectronic printing; the conductive layer includes a plurality of discrete conductive blocks, and the conductive blocks cover a group of the through holes, the first point and the second point; A sensitive layer is formed on a side of the flexible layer facing away from the substrate; the sensitive layer includes a plurality of discrete film blocks, and the film blocks at least partially overlap with the through holes.

2. The preparation method according to claim 1, characterized in that, The flexible layer comprises a polydimethylsiloxane film layer; the method of forming the flexible layer on the second surface by microelectronic printing comprises: The prepolymer and the curing agent are mixed evenly at a mass ratio of M:1 to obtain a mixed adhesive, and the mixed adhesive is vacuumed to remove bubbles; wherein 8≤M≤10; The mixed glue is used as a raw material, and the polydimethylsiloxane film layer is printed by a dispensing method.

3. The preparation method according to claim 2, wherein, The method of printing the polydimethylsiloxane film layer by dispensing includes: Based on a first printing speed and a first printing pressure, the polydimethylsiloxane film layer is printed by dispensing; wherein, the first printing speed is: 5-15 mm / s, and the first printing pressure is: 30-90 KPa.

4. The preparation method according to claim 2 or 3, characterized in that, After printing the polydimethylsiloxane film layer by dispensing, the preparation method further comprises: The substrate is heated at a first heating temperature until the polydimethylsiloxane film layer is solidified; wherein the first heating temperature is 60-80°C.

5. The preparation method according to claim 1, characterized in that, The conductive layer includes a silver paste layer; the forming of the conductive layer on the first surface by microelectronic printing includes: The conductive blocks are printed by using conductive silver paste as a raw material and a dispensing method.

6. The preparation method according to claim 5, characterized in that, The method of printing the conductive block by dispensing comprises: Based on the second printing speed and the second printing pressure, the conductive block is printed by dispensing; wherein the second printing speed is 4 to 15 mm / s, and the second printing pressure is 70 to 100 KPa.

7. The preparation method according to claim 5 or 6, characterized in that, After printing the conductive block in a dispensing manner, the preparation method further comprises: The substrate is heated at a second heating temperature until the conductive block is solidified; wherein the second heating temperature is 80 to 120° C. and the heating time is 30 to 50 minutes.

8. The preparation method according to claim 1, characterized in that, The sensitive layer comprises a nano-gold layer; and the sensitive layer is formed on a side of the flexible layer away from the substrate, comprising: The thin film block is formed on a side of the flexible layer facing away from the substrate by magnetron sputtering.

9. The preparation method according to claim 8, characterized in that, The method of forming the thin film block on a side of the flexible layer away from the substrate by magnetron sputtering comprises: Based on a first sputtering time, the thin film block is sputtered on a side of the flexible layer away from the substrate; wherein the first sputtering time is 1 to 2 minutes.

10. The preparation method according to claim 8 or 9, characterized in that, Also includes: Modify an antibody on a side of the thin film block facing away from the flexible layer; The antibody is used for specifically binding to a biomolecule to be detected.

11. A biochemical sensing array, characterized in that, The biochemical sensing array is formed based on the preparation method according to any one of claims 1 to 10.

12. The biochemical sensing array according to claim 11, wherein A signal interface is further provided on a first surface of the substrate. The signal interface includes a first signal interface and a second signal interface arranged in sequence. The first signal interface is electrically connected to the first trace, and the second signal interface is electrically connected to the second trace in one-to-one correspondence.

13. The biochemical sensing array according to claim 12, wherein The diameter of the through hole is: 2.0 to 5.0 mm; The distance between the centers of adjacent through holes is: 3.0 to 15 mm; The distance between the edge of the first point position and the edge of the corresponding through hole is: 0.3 to 1.0 mm; The distance between the edge of the second point position and the edge of the corresponding through hole is: 0.3 to 1.0 mm; The line width of the first trace is: 0.2 to 0.5 mm; The line width of the second trace is: 0.2 to 0.5 mm; The line pitch between the first trace and the second trace and between the second traces is: 0.2 to 0.5 mm; The width of the first signal interface is: 0.3 to 0.7 mm; The width of the second signal interface is: 0.3 to 0.7 mm.

14. A test strip, characterized in that, Comprising: The biochemical sensing array according to any one of claims 11 to 13.

15. A detection kit, characterized in that, Comprising: The test strip according to claim 14.

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